Evolutionary loss of inflammasomes in the Carnivora and implications for the carriage of zoonotic infections.

Zoonotic pathogens, such as COVID-19, reside in animal hosts before jumping species to infect humans. The Carnivora, like mink, carry many zoonoses, yet how diversity in host immune genes across species affect pathogen carriage is poorly understood. Here, we describe a progressive evolutionary downregulation of pathogen-sensing inflammasome pathways in Carnivora. This includes the loss of nucleotide-oligomerization domain leucine-rich repeat receptors (NLRs), acquisition of a unique caspase-1/-4 effector fusion protein that processes gasdermin D pore formation without inducing rapid lytic cell death, and the formation of a caspase-8 containing inflammasome that inefficiently processes interleukin-1ß. Inflammasomes regulate gut immunity, but the carnivorous diet has antimicrobial properties that could compensate for the loss of these immune pathways. We speculate that the consequences of systemic inflammasome downregulation, however, can impair host sensing of specific pathogens such that they can reside undetected in the Carnivora.

Truncation of TRIM5 in the Feliformia explains the absence of retroviral restriction in cells of the domestic cat.

TRIM5alpha mediates a potent retroviral restriction phenotype in diverse mammalian species. Here, we identify a TRIM5 transcript in cat cells with a truncated B30.2 capsid binding domain and ablated restrictive function which, remarkably, is conserved across the Feliformia. Cat TRIM5 displayed no restriction activity, but ectopic expression conferred a dominant negative effect against human TRIM5alpha. Our findings explain the absence of retroviral restriction in cat cells and suggest that disruption of the TRIM5 locus has arisen independently at least twice in the Carnivora, with implications concerning the evolution of the host and pathogen in this taxon.

Changes in the fecal concentrations of cortisol and androgen metabolites in captive male jaguars (Panthera onca) in response to stress

In the present study we determined the efficacy of the measurement of fecal cortisol and androgen metabolite concentrations to monitor adrenal and testicular activity in the jaguar (Panthera onca). Three captive male jaguars were chemically restrained and electroejaculated once or twice within a period of two months. Fecal samples were collected daily for 5 days before and 5 days after the procedure and stored at -20ºC until extraction. Variations in the concentrations of cortisol and androgen metabolites before and after the procedure were determined by solid phase cortisol and testosterone radioimmunoassay and feces dry weight was determined by drying at 37ºC for 24 h under vacuum. On four occasions, fecal cortisol metabolite levels were elevated above baseline (307.8 ± 17.5 ng/g dry feces) in the first fecal sample collected after the procedure (100 to 350 percent above baseline). On one occasion, we did not detect any variation. Mean (± SEM) fecal androgen concentration did not change after chemical restraint and electroejaculation (before: 131.1 ± 26.7, after: 213.7 ± 43.6 ng/g dry feces). These data show that determination of fecal cortisol and androgen metabolites can be very useful for a noninvasive assessment of animal well-being and as a complement to behavioral, physiological, and pathological studies. It can also be useful for the study of the relationship between adrenal activity and reproductive performance in the jaguar.

Noninvasive monitoring of adrenocortical activity in carnivores by fecal glucocorticoid analyses.

Measurement of glucocorticoid metabolites in feces has become an accepted method for the noninvasive evaluation of adrenocortical activity. The objective of this study was to determine if a simple cortisol enzyme immunoassay (EIA) was suitable for monitoring adrenocortical activity in a variety of carnivore species. Performance of the cortisol EIA was gauged by comparison to a corticosterone radioimmunoassay (RIA) that has been used for measuring glucocorticoid metabolites in feces of numerous species. Tests for parallelism and extraction efficiency were used to compare the cortisol EIA and corticosterone RIA across eight species of carnivores (Himalayan black bear, sloth bear, domestic cat, cheetah, clouded leopard, black-footed ferret, slender-tailed meerkat, and red wolf). The biological relevance of immunoreactive glucocorticoid metabolites in feces was established for at least one species of each Carnivora family studied with an adrenocorticotropic hormone (ACTH) challenge. High performance liquid chromatography (HPLC) analysis of fecal extracts for each species revealed (1) the presence of multiple immunoreactive glucocorticoid metabolites in feces, but (2) the two immunoassays measured different metabolites, and (3) there were differences across species in the number and polarities of metabolites identified between assay systems. ACTH challenge studies revealed increases in fecal metabolite concentrations measured by the cortisol EIA and corticosterone RIA of approximately 228-1145% and approximately 231-4150% above pre-treatment baseline, respectively, within 1-2 days of injection. Concentrations of fecal glucocorticoid metabolites measured by the cortisol EIA and corticosterone RIA during longitudinal evaluation (i.e., >50 days) of several species were significantly correlated (P<0.0025, correlation coefficient range 0.383-0.975). Adrenocortical responses to physical and psychological stressors during longitudinal evaluations varied with the type of stimulus, between episodes of the same stimulus, and among species. Significant elevations of glucocorticoid metabolites were observed following some potentially stressful situations [anesthesia (2 of 3 subjects), restraint and saline injection (2 of 2 subjects), restraint and blood sampling (2 of 6 episodes), medical treatment (1 of 1 subject)], but not in all cases [e.g., gonadotropin injection (n=4), physical restraint only (n=1), mate introduction/breeding (n=1), social tension (n=1), construction (n=2) or relocation (n=1)]. Results reinforced the importance of an adequate baseline period of fecal sampling and frequent collections to assess adrenocortical status. The corticosterone RIA detected greater adrenocortical responses to exogenous ACTH and stressful exogenous stimuli in the Himalayan black bear, domestic cat (female), cheetah, clouded leopard, slender-tailed meerkat, and red wolf, whereas the cortisol EIA proved superior to resolving adrenocortical responses in the black-footed ferret and domestic cat (male). Overall results suggest the cortisol EIA tested in this study offers a practical method for laboratories restricted in the usage of radioisotopes (e.g., zoological institutions and field facilities) to integrate noninvasive monitoring of adrenocortical activity into studies of carnivore behavior and physiology.

Continuous melatonin treatment and fasting in the raccoon dog (Nyctereutes procyonoides)--vernal body weight regulation and reproduction.

The raccoon dog (Nyctereutes procyonoides) is a canid omnivore with marked seasonal changes in its body adiposity. The aim of this study was to investigate the roles of melatonin, leptin, ghrelin and growth hormone (GH) in weight regulation and reproduction of the species. Sixteen raccoon dogs were treated with continuous-release melatonin implants in Aug 2000 and in Feb 2001 (the MEL group) and 16 animals were sham-operated (the SHAM group). Half of the raccoon dogs were fasted between Nov 27(th) 2000 and Jan 25(th) 2001. The autumnal results have been previously published and this paper reports the vernal data. The leptin concentrations of the SHAM females were high before the mating season, decreased before estrus, increased during gestation and reduced after parturition. The MEL females had higher leptin concentrations than the SHAM females in early March, whereas the MEL males had lower leptin concentrations than the SHAM males in late March. Also the ghrelin and GH concentrations of the SHAM females decreased before estrus. Continuous melatonin treatment advanced the vernal rise in the ghrelin concentrations and the vernal drop and the subsequent rise in the GH concentrations of the females. Melatonin also increased their body mass indices from July to Aug 2001, indicating that it triggers the autumnal accumulation of fat in the species.

Effects of phytosterols on the endocrinology and metabolism of the female raccoon dog (Nyctereutes procyonoides).

Phytosterols (PS) are the analogues of animal cholesterol in various plants. beta-Sitosterol is a PS used in margarines and natural remedies to lower elevated serum cholesterol levels. PS enter the ecosystem via pulp mill effluents. The study investigated the endocrine and metabolic effects of PS on the female raccoon dog (Nyctereutes procyonoides), a canid omnivore. Eight female animals were exposed perorally to 8 mg PS/kg/d for 4 wk with 8 animals in the control group. In the PS-treated females, there was a transitory decrease in the plasma estradiol concentrations with an increase in the plasma follicle-stimulating hormone levels. The plasma triiodothyronine concentrations were higher in the PS group. Serum lipid concentrations decreased in PS-treated and control animals. This probably represents a seasonal adaptation. Most of the cholesterol in raccoon dog serum was high-density lipoprotein cholesterol, unlike that in humans but similar to some other carnivores. Liver and kidney ethoxyresorufin O-deethylase activities were lower in the PS treated females. Data indicate that raccoon dogs may not be a sentinel species for PS effects.

Phytosterols act as endocrine and metabolic disruptors in the European polecat (Mustela putorius).

Phytosterols or plant sterols (PS) are consumed as natural remedies and margarines by the general population in developed countries to lower elevated serum cholesterol levels. They are also present in high concentrations in pulp mill effluents. The aim of the study was to screen the endocrine and metabolic parameters of the European polecat (Mustela putorius) for the effects of PS. The results showed an increase in the plasma estradiol and TH levels with no effects on the hypophyseal regulatory hormones. The plasma ghrelin levels decreased. PS also affected intermediary metabolism. The liver glycogen content increased as did the kidney glucose-6-phosphatase activity. The liver lipase esterase activity, on the other hand, decreased due to PS. In serum lipids the total cholesterol did not change, but the low-density lipoprotein levels increased and the high-density lipoprotein-cholesterol ratio decreased. PS had widespread previously unreported effects on the physiology of the polecat. The multiple effects indicate the need of a thorough risk assessment of the effects and interactions of PS.

Noninvasive fecal monitoring of glucocorticoids in spotted hyenas, Crocuta crocuta.

The aim of this study was to validate a method for measuring glucocorticoids noninvasively in feces of spotted hyenas (Crocuta crocuta). Three established enzyme immunoassays (EIA) for cortisol, corticosterone, and 11-oxoetiocholanolone were tested, but proved unsatisfactory. A new EIA using another corticosterone antibody was established and was used for all subsequent analyses; this EIA was validated by demonstrating parallelism between serial dilutions of spotted hyena fecal extracts and dilutions of standard corticosterone and by the recovery of corticosterone added to fecal extracts. High-performance liquid chromatography (HPLC) fractions analyzed by EIA showed various immunoreactive substances with polarities of unconjugated steroids. The physiological relevance of fecal glucocorticoid metabolites was further validated by demonstrating that (1) injection of exogenous ACTH to four males and two females led to a significant increase in fecal glucocorticoid metabolites within 24-50 h, (2) the translocation of a male spotted hyena to a new enclosure resulted in a fivefold increase compared to baseline concentrations, and (3) agonistic social interactions and physical conflict resulted in large increases of fecal glucocorticoid metabolites in both protagonists. Fecal steroid assessment is therefore of use in monitoring adrenal activity in spotted hyenas.

Comparative binding affinity study of progestins to the cytosol progestin receptor of endometrium in different mammals.

The relative binding affinity of 5 alpha-reduced progestins and a newly synthesized antiprogestin J912 (progesterone 100%) was determined in a competitive receptor binding assay using [3H]ORG-2058 as radiolabeled ligand for the progestin receptor. Uteri obtained from 12 different species of four mammalian orders were examined. The relative binding affinity of 75-100% and a blood prevalence of 5 alpha-pregnane-3,20-dione in horses and African elephants suggest a biological role of this particular 5 alpha-reduced progesterone. For pigs the binding affinity of 5 alpha-pregnane-3,20-dione was about 50% of progesterone, but blood levels are unknown. In all other cases the low binding affinity of investigated progestins precludes possible biological role. For 5 alpha-pregnane-3 alpha-ol-20-one, 5 alpha-pregnane-20 alpha-ol-3-one, and 5 alpha-pregnane-3 beta,20 alpha-diol the relative binding affinity was less than 1%. A rather low binding (< 15%) was observed in 5 alpha-pregnane-3,20-dione in all ruminant species investigated. The antiprogestin J912 was found to be highly efficient in displacing progesterone from its endometrial binding sites in carnivores and might therefore be used for pregnancy interruption during diapause in certain species, e.g., in captive bears.

Excretion profiles of some reproductive steroids in the faeces of captive Nepalese red panda (Ailurus fulgens fulgens).

Faecal samples were collected up to once daily from three female Nepalese red panda (Ailurus fulgens fulgens) prior to, during, and after the expected 1995 breeding season. Radioimmunoassay of faecal progestins and oestrogens showed hormone profiles that suggest that this species is a seasonally polyoestrous, induced ovulator. Examination of faecal extracts following high pressure liquid chromatography and using antisera of high specificity, demonstrated at least five faecal progestins but only one major oestrogen, probably oestradiol. Samples from males were collected up to once weekly during the breeding season in 1996, and androgens measured by radioimmunoassay. Faecal androgens fluctuated widely for all males.

Felinine: a urinary amino acid of Felidae.

Felinine (2-amino-7-hydroxy-5,5-dimethyl-4-thiaheptanoic acid) has been identified in the urine of several members of the Felidae family including the cat (Felis catus). Rates of excretion of 95 mg/day have been recorded for entire male cats with entire female cats excreting around 19 mg/day. These high excretion rates in entire male cats may have a significant effect on the daily sulphur amino acid requirement. The isoamyl moiety of felinine seems to originate from the same isoprenoid pool as used for the synthesis of cholesterol in the cat. The sulphur in the felinine molecule appears to originate from cysteine, although some contradictory evidence exists. The site of synthesis and the method of transportation in the blood remain largely unknown. The biological significance of felinine to the animal is still a matter for speculation, but its function as a precursor to a pheromone seems likely. Recently, an accurate chemical assay for felinine has been developed that will allow investigation of felinine in different tissues and excretions and from a wider range of mammals.

Reproductive impairment in the Florida panther: nature or nurture?

Many of the remaining members of the endangered Florida panther (Felis concolor coryi) population suffer from one or more of a variety of physiological, reproductive, endocrine, and immune system defects including congenital heart defects, abnormal sperm, low sperm density, cryptorchidism, thyroid dysfunction, and possible immunosuppression. Mercury contamination, determined to be the cause of death of a female panther in 1989, was presented as the likely cause of thyroid dysfunction. As genetic diversity in the species was less than expected, all of the other abnormalities have been attributed to inbreeding. However, exposure to a variety of chemical compounds, especially those that have been identified as environmental endocrine disrupters (including mercury, p,p'-DDE, and polychlorinated biphenyls), has elicited all of the listed abnormalities in other species. A number of these contaminants are present in South Florida. An exposure pathway has been identified, and evidence presented in this paper, including the fact that there appears to be no significant difference between serum estradiol levels in males and females, suggests that many male panthers may have been demasculinized and feminized as a result of either prenatal or postnatal exposure. Thus, regardless of the effects of inbreeding, current evidence seems to indicate that environmental contaminants may be a major factor contributing to reproductive impairment in the Florida panther population.

A carnivore species (Canis familiaris) expresses circadian melatonin rhythm in the peripheral blood and melatonin receptors in the brain.

Dogs kept under controlled photoperiodic conditions of 12 h light and 12 h dark expressed a clear diurnal melatonin rhythm in the peripheral blood, with a swift peak restricted to the late part of the scotophase. The highest density of high-affinity, G-protein-linked 2-[125I]iodomelatonin binding sites was found in the pars tuberalis of the pituitary gland. Binding sites were found also in the pars distalis, and light microscopy/high-resolution autoradiography showed that binding was located exclusively over the chromophobe and basophilic cells forming the adenopituitary zona tuberalis, well developed in this species, and extending into the gland as a continuation of pars tuberalis. Cords of basophilic cells located in the pars distalis proper also expressed high receptor density. The eosinophils in the adenohypophysis and the neural lobe were devoid of binding. Heavily labeled were the external laminar and the mitral cell layers of the olfactory bulbs, but no binding was detected in the filae nervi olfactorii or tractus olfactorius. The hypothalamic suprachiasmatic nuclei were discernible clearly. Quantitative autoradiography inhibition experiments revealed that the apparent melatonin inhibitory constant (IC50) in all those areas was around 0.1 nmol/l, which is a physiologically appropriate value considering the peripheral blood melatonin levels. Co-incubation with guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) led to a consequential decrease in the binding density. The specific binding observed in other areas (hippocampus, frontal, parietal, occipital cortex and cerebellum) was rather weak, diffuse and could not be attributed to a particular layer; the apparent IC50 for melatonin was about 1 mumol/l, and co-incubation with GTP gamma S did not modify the binding density. Collectively, these data show that the dog possess all the prerequisites for an efficient network adapted to photoperiodic time measurements. A circadian melatonin signal in the peripheral blood and an apparently functional readout receptor system located in key positions within the brain are both present in this species.

A mechanism for virilization of female spotted hyenas in utero.

Female spotted hyenas exhibit male-like genitalia and dominance over males. Hyena ovarian tissues incubated in vitro produced large quantities of the steroid hormone precursor androstenedione. The activity of aromatase, which converts androstenedione to estrogen, was one-twentieth as great in hyena versus human placental homogenates. In comparison, the activity of 17 beta-hydroxysteroid dehydrogenase, which converts androstenedione to testosterone, was equal in the two homogenates. The limited aromatase activity may allow the hyena placenta to convert high circulating concentrations of androstenedione to testosterone, which results in virilization of the fetal external genitalia and possibly destruction of fetal ovarian follicles. Androstenedione production by residual ovarian stromal cells during reproductive life accounts for the epigenetic transmission of virilization in female spotted hyenas.

[Clinical experience and comparison of ketamine-medetomidine with ketamine-xylazine anesthesia in the African wild dog (Lycaon pictus) in captivity]. / Klinické zkusenosti a srovnání ketamín-medetomidin a ketamín-xylazín anestézie psa hyenového (Lycaon pictus) v zajetí.

The effects of two mixtures, ketamine-xylazine and ketamine-medetomidine, were compared in anesthesia of African wild dog (Lycaon pictus) in the Zoo park at Dvur Králové; the effects of these combinations were also investigated on the triad values and on the basic hematological (red blood counts, hematocrit, hemoglobin content, derived parameters MCM, MCHC, MCV, white blood counts, differential blood counting) and biochemical (total proteins, glucose, creatinine, urea, cholesterol, magnesium, calcium, phosphorus, chlorides, sodium, potassium, alkaline phosphatase, gamma-glutamyl transferase, alanine transferase, aspartate transaminase) parameters of blood and blood serum in anesthesia. Tab. I shows the reasons for anesthesia in 68 individuals of African wild dog in the years 1980-1990. As for both mixtures, application of drugs with a blowpipe was used. In the anesthetized animals, the outset of ataxia, laying down and the outset of sleep were followed (Tab. II), as well as the time of wakening up without and with antidote administration (Tabs. IV and V). The outset of ataxia was fast 1.5 +/- 0.6 min) in the ketamine-xylazine mixture administered at doses of 5.07 +/- 1.16 mg/kg ketamine and 2.11 +/- 0.53 mg/kg xylazine, similarly like lying down (3.2 +/- 1.0 min) and loosing sensation (6.3 +/- 1.6 min). At the start of the drug action, vomiting was often observed, and sometimes in the first ten minutes after drug harpooning short clonic convulsions of the limbs or the whole body occurred. In further course, immobilization and anesthesia were complete and satisfactory in all cases. The first reactions to outer stimuli during wakening up without antidote administration were observed in 135 +/- 11.9 minutes while the animals stood up in 210 +/- 44.5 minutes after drug harpooning (Tab. IV). When the nonspecific antidote xylazine-yohimbine was used, the first reactions after i.m. instillation at a dose of 0.31 +/- 0.02 mg/kg appeared in 20.3 +/- 0.57 min, after i.v. instillation at a dose of 0.11 +/- 0.17 it was in 9.3 +/- 4.16 min after antidote administration. The animals stood up and started walking in 53.3 +/- 24.3 min after i.v. instillation (Tab. V). No significant changes (Tab. III) were observed in the values of temperature, breathing rate and pulse rate when the variations of triad values were investigated in five individuals within the first 30 minutes (up to 10 min, 20 and 30 min) after drug harpooning.(ABSTRACT TRUNCATED AT 400 WORDS)

Measurements of faecal oestradiol and progesterone in non-pregnant and pregnant domestic and exotic cats.

Faecal samples collected for variable periods from 12 animals and five species of cats were assayed for progesterone and oestradiol content by application of standard radioimmunoassays to aliquots (50 microliters) of methanol extracts (4 ml) of a mixture of 0.5 g sample, 0.5 ml water and 1 g aluminium oxide, following partitioning of the total extract with petroleum ether (3 ml), further dilution of assay aliquots and drying. Recoveries averaged 100 and 72% for oestradiol and progesterone, respectively. Results included increases in progesterone during luteal phases or pregnancies to 7688 ng g-1 (tiger, Panthera tigris), 2594 ng g-1 (lion, P. leo), 3000 ng g-1 (cheetah, Acinonyx jubatus) and 4915 ng g-1 (caracal, Felis caracal). Faecal oestradiol peaks near oestrus included 246 ng g-1 (tiger), 175 ng g-1 (lion) 190 ng g-1 (cheetah), 23 ng g-1 (caracal) and 190 ng g-1 (domestic cat, F. catus).

Sex-specific androgen binding in spotted hyaena (Crocuta crocuta) plasma.

1. A charcoal adsorption assay demonstrated a large variance in androgen binding ability in female spotted hyaenas. 2. A positive correlation between plasma androgen binding ability and ovarian steroid concentrations was demonstrated in adult females. 3. The strong plasma binding affinity for testosterone and dihydrotestosterone (DHT) (nM) together with the lack of cortisol and weaker oestradiol-17 beta binding suggests that a specific androgen binding substance, possibly a protein, is present in adult females of this species. 4. The lack of high affinity binding in male spotted hyaenas is unusual and deserves further investigation. 5. Some androgen binding in all, including males and immature animals suggests that albumin may bind some plasma androgens in this species.

Hormonal correlates of 'masculinization' in female spotted hyaenas (Crocuta crocuta). 2. Maternal and fetal steroids.

Concentrations of androgens (androstenedione, testosterone, 5 alpha-dihydrotestosterone), oestrogen and progesterone were measured in relation to pregnancy in the spotted hyaena (Crocuta crocuta). The gestation period was estimated to be about 110 days. There was a marked progressive rise in all the steroids starting in the first third of gestation. Chromatographic separation of plasma showed that much of the oestrogen is not oestradiol (only 12% of total measured) and that a significant fraction of the 'testosterone' may be dihydrotestosterone. In the final third of pregnancy, concentrations of androgen (especially testosterone plus dihydrotestosterone) in the female circulation reached the maximal values of adult males; the percentage of dihydrotestosterone relative to total testosterone plus dihydrotestosterone was higher in females (44 +/- 3.9%, n = 20) than in males (29.5 +/- 3.5%, n = 17). Plasma androstenedione was also significantly higher in females, but the increment was less than for oestrogen, testosterone and progesterone, and the temporal pattern was less clear. Samples from the maternal uterine and ovarian circulation showed that androstenedione is largely of ovarian origin and metabolized by the placenta, while testosterone, progesterone and oestrogen are primarily of placental or uterine origin. Fetal samples were taken from two mixed-sex sets of twins and one male singleton. Gradients across the placenta measured in the fetal circulation confirmed that the placenta metabolizes androstenedione and is a source of testosterone for the female fetus; there were no consistent differences in androgens between male and female fetuses. It is suggested that the conspicuous masculinization of the female spotted hyaena, especially evident in the external genitalia at birth, is a result, at least in part, of high placental production of testosterone or dihydrotestosterone derived from the metabolism of high maternal androstenedione.

[Histological variants of the hepatoid glands in representative Canidae and Cavicornia: the structural and histochemical characteristics]. / Gistologicheskie varianty gepatoidnykh zhelez u predstavitelei volch'ikh i polorogikh: strukturnye i gistokhimicheskie kharakteristiki.

Both the original material and the data from literature collected in two previous papers, show a considerable diversity of structure and secretion of the glands which constitute the hepatoid gland type. According to the body of available histological (including morphometrical) and histochemical data, the authors distinguish: a) features characteristic of the hepatoid gland type only which do not appear in other types of skin glands; b) seven histological varieties within the hepatoid gland type. These seven varieties can be united, according to the laws of their structure and function, into two groups (1-3 and 5-7), with an intermediate variety (4) between them. The first group includes hepatoid glands of the "classical" structure which has been previously detaily exampled by circumanal glands of the dog: these glands possess cysts, produce a purely protein secretion; the secretion type is merocrine. The second group comprises mero-holocrine hepatoid glands which are characterized by the absence of cysts, broader excretion ducts, and a capability to produce, besides protein, considerable amounts of melanin and/or hydrophobic lipids.

Amyloid in the pancreatic islets of the cougar (Felis concolor) is derived from islet amyloid polypeptide (IAPP).

1. Islet amyloid isolated from the pancreas of a 20-year-old cougar (Felis concolor) was dissolved and purified by gel permeation and reversed phase HPLC for amino acid sequence analysis. 2. N-Terminal amino acid sequence analysis of the purified protein revealed a primary structure (positions 1-28) identical to islet amyloid polypeptide (IAPP) from domesticated cats. 3. IAPP from the cougar, like IAPP from the human and domesticated cat, incorporates an inherently amyloidogenic AILS sequence at positions 25-28.